Figure 5

BMS-345541 or wortmannin inhibited LPS-induced nuclear translocation of phospho p65 in chondrocytes as demonstrated by immunofluorescence microscopy. Primary human chondrocyte cultures either served as controls (a-b: without primary antibody; c-d: with primary antibody) or were treated with lipopolysaccharides (LPS) alone (e-f) or were pre-treated with wortmannin (20 nM) (g-h) or with BMS-345541 (5 mM) (i-j) for 12 h before co-treatment with LPS (100 ng/ml) for 24 h before immunolabeling with phospho p65 antibodies and rhodamine-coupled secondary antibodies and counterstained with DAPI to visualize cell nuclei. Images shown are representative of three independent experiments. Magnification x400; bar, 30 nm.